Purification and amino acid sequences of six Tx3 type neurotoxins from the venom of the Brazilian ‘armed’ spider Phoneutria Nigriventer (keys.)

Cordeiro, Marta do Nascimento; Figueiredo, Suely G.; Valentim, Ana do Carmo; Diniz, Carlos R.; Eickstedt, V.R.D.; Gilroy, John; Richardson, Michael

doi:10.1016/0041-0101(93)90354-l

Cited by 143 publications

(78 citation statements)

References 16 publications

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“…Tx3-4 was purified as described by Cordeiro et al [19], dissolved in water (80 µM) and stored in aliquots at k20 mC. -[$H]Glutamate was purchased from Amersham International (Little Chalfont, Bucks., U.K.).…”

Section: Experimental Materialsmentioning

confidence: 99%

Inhibition of glutamate uptake by a polypeptide toxin (phoneutriatoxin 3-4) from the spider Phoneutria nigriventer

Reis

Prado

Kalapothakis

et al. 1999

Biochemical Journal

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Glutamate concentration increases significantly in the extracellular compartment during brain ischaemia and anoxia. This increase has an important Ca2+-independent component, which is due in part to the reversal of glutamate transporters of the plasma membrane of neurons and glia. The toxin phoneutriatoxin 3-4 (Tx3-4) from the spider Phoneutria nigriventer has been reported to decrease the evoked glutamate release from synaptosomes by inhibiting Ca2+ entry via voltage-dependent Ca2+ channels. However, we report here that Tx3-4 is also able to inhibit the uptake of glutamate by synaptosomes in a time-dependent manner and that this inhibition in turn leads to a decrease in the Ca2+-independent release of glutamate. No other polypeptide toxin so far described has this effect. Our results suggest that Tx3-4 can be a valuable tool in the investigation of function and dysfunction of glutamatergic neurotransmission in diseases such as ischaemia.

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Section: Experimental Materialsmentioning

confidence: 99%

Inhibition of glutamate uptake by a polypeptide toxin (phoneutriatoxin 3-4) from the spider Phoneutria nigriventer

Reis

Prado

Kalapothakis

et al. 1999

Biochemical Journal

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“…P. nigriventer venom was obtained by electrical stimulation of anesthetized spiders. Venom was centrifuged at 4000g for 10 min, and the supernatant was fractionated by gel filtration on columns of Sephadex G-50 superfine and superose 12HR, and reverse phase fast protein liquid chromatography on C 2 /C 8 (PEP-RPC) and C 1 /C 8 (PRO-RPC) columns as described in detail previously (Rezende et al, 1991;Cordeiro et al, 1993). Peptides were detected by monitoring the absorbance at 216 nm (Cordeiro et al, 1993).…”

Section: Methodsmentioning

confidence: 99%

“…Venom was centrifuged at 4000g for 10 min, and the supernatant was fractionated by gel filtration on columns of Sephadex G-50 superfine and superose 12HR, and reverse phase fast protein liquid chromatography on C 2 /C 8 (PEP-RPC) and C 1 /C 8 (PRO-RPC) columns as described in detail previously (Rezende et al, 1991;Cordeiro et al, 1993). Peptides were detected by monitoring the absorbance at 216 nm (Cordeiro et al, 1993). The fraction PhTx3 obtained from the chromatography on a PRO-RPC column (Rezende et al, 1991) was dissolved in 1 ml of 0.1% (v/v) aqueous trifluoroacetic acid (TFA) and subjected to reverse phase HPLC on a preparative column (22 mm ϫ 25 cm) of Vydac C18 (218TP1022; Technicol Ltd., Stockport, UK) equilibrated in the same solvent.…”

Section: Methodsmentioning

confidence: 99%

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Inhibition of High Voltage-Activated Calcium Channels by Spider Toxin PnTx3-6

Vieira¹,

Kushmerick²,

Hildebrand³

et al. 2005

J Pharmacol Exp Ther

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104

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Animal peptide toxins have become powerful tools to study structure-function relationships and physiological roles of voltage-activated Ca 2ϩ channels. In the present study, we investigated the effects of PnTx3-6, a neurotoxin purified from the venom of the spider Phoneutria nigriventer on cloned mammalian Ca 2ϩ channels expressed in human embryonic kidney 293 cells and endogenous Ca 2ϩ channels in N18 neuroblastoma cells. Whole-cell patch-clamp measurements indicate that PnTx3-6 reversibly inhibited L-(␣ 1C

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“…Such neurotoxins are acylpolyamines, proteins, and peptides in nature. The venoms from Agelenopsis aperta of the American (USA) subcontinent [8][9][10][11] and Phoneutria nigriventer of the Brazilian subcontinent [12,13] are the most explored species for neurotoxicity among the spider venoms.…”

Section: Introductionmentioning

confidence: 99%

Purification and Characterization of a Nonenzymatic Neurotoxin fromHippasa partita(Lycosidae) Spider Venom Gland Extract

Nagaraju

Kemparaju

2013

Journal of Toxins

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India is a habitat for nearly one thousand four hundred forty-seven species of spiders under three hundred and sixty-five genera and sixty families. Our initial survey on toxic bite by spider revealed severe edema, itching, acute pain, and hemorrhage following tissue necrosis, which are the general symptoms of envenomation, but there are no reports of mortality. Significantly,Hippasa partitaspider, commonly called “funnel web spider,” which is endemic in hilly regions of the Western Ghats is responsible for envenomation. In this study, a nonenzymatic neurotoxin has been purified fromH. partitavenom gland extract. Gel filtration and ion exchange chromatography were used to purify the toxin into homogeneity as shown by SDS-PAGE, RP-HPLC, and MALDI-TOF. Neurotoxin is devoid of enzymatic activities but causes intense neurotoxic symptoms. Neurotoxin is found to inhibit the twitch response of sciatic nerve gastrocnemius muscle preparation and is found to be postsynaptic in action. Neurotoxin is devoid of coagulant activity, edema, and hemorrhage and is nonlethal to mice (up to 5 mg/kg body weight). In conclusion, a neurotoxin, which is a principle agent in whole venom responsible for induced neurotoxic symptoms, has been purified and characterized.

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Purification and amino acid sequences of six Tx3 type neurotoxins from the venom of the Brazilian ‘armed’ spider Phoneutria Nigriventer (keys.)

Cited by 143 publications

References 16 publications

Inhibition of glutamate uptake by a polypeptide toxin (phoneutriatoxin 3-4) from the spider Phoneutria nigriventer

Inhibition of glutamate uptake by a polypeptide toxin (phoneutriatoxin 3-4) from the spider Phoneutria nigriventer

Inhibition of High Voltage-Activated Calcium Channels by Spider Toxin PnTx3-6

Purification and Characterization of a Nonenzymatic Neurotoxin fromHippasa partita(Lycosidae) Spider Venom Gland Extract

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